Photo-induced Proton Transfers of Microbial Rhodopsins

Molecular Photochemistry - Various Aspect

Succession of Genetic Diversity of Botryococcus braunii (Trebouxiophyceae) in Two Japanese Reservoirs

AbstractBotryococcus braunii is a green colonial alga that produces large volumes of liquid hydrocarbon. Therefore, B. braunii is expected to be useful as an alternative fuel resource. Natural blooms of B. braunii have been recorded in several lakes and reservoirs. Elucidation of natural B. braunii blooming would provide important information for the development of an open-pond cultivation system. In this study, we periodically assessed the genetic diversity and colony density of B. braunii populations, along with several environmental parameters, in two Japanese reservoirs (provisionally called “N” and “S”) from December 2008 to December 2009. Reservoir N had low numbers of B. braunii colonies whereas Reservoir S was characterized by periodic density increases that occurred in December 2008, and in March, September, and December 2009. Population genetics analysis using specific environmental sequences (PGA-SES method) was conducted for B. braunii populations for the first time. Among the B. braunii-dominated samples of Reservoir S, high levels of genetic diversity were observed in December 2008 and March 2009, whereas the diversity levels in September and December 2009 were low. The results suggest that B. braunii periodicity can be categorized into a high genetic diversity type and a low genetic diversity type. The high genetic diversity type may be caused by simultaneous growth of many genotypes, whereas the low genetic diversity type seems to be explained by increases in the cell density of only a few adapted genotypes

Desiccation tolerance of Botryococcus braunii (Trebouxiophyceae, Chlorophyta) and extreme temperature tolerance of dehydrated cells

Botryococcus braunii Kützing, a green colonial microalga, occurs worldwide in both freshwater and brackish water environments. Despite considerable attention to B. braunii as a potential source of renewable fuel, many ecophysiological properties of this alga remain unknown. Here, we examined the desiccation and temperature tolerances of B. braunii using two newly isolated strains BOD-NG17 and BOD-GJ2. Both strains survived through 6- and 8-month desiccation treatments but not through a 12-month treatment. Interestingly, the desiccation-treated cells of B. braunii gained tolerance to extreme temperature shifts, i.e., high temperature (40 °C) and freezing (−20 °C). Both strains survived for at least 4 and 10 days at 40 and −20 °C, respectively, while the untreated cells barely survived at these temperatures. These traits would enable long-distance dispersal of B. braunii cells and may account for the worldwide distribution of this algal species. Extracellular substances such as polysaccharides and hydrocarbons seem to confer the desiccation tolerance

Photochemical characterization of actinorhodopsin and its functional existence in the natural host

Actinorhodopsin (ActR) is a light-driven outward H+ pump. Although the genes of ActRs are widely spread among freshwater bacterioplankton, there are no prior data on their functional expression in native cell membranes. Here, we demonstrate ActR phototrophy in the native actinobacterium. Genome analysis showed that Candidatus Rhodoluna planktonica, a freshwater actinobacterium, encodes one microbial rhodopsin (RpActR) belonging to the ActR family. Reflecting the functional expression of RpActR, illumination induced the acidification of the actinobacterial cell suspension and then elevated the ATP content inside the cells. The photochemistry of RpActR was also examined using heterologously expressed RpActR in Escherichia coli membranes. The purified RpActR showed lambda(max) at 534 nm and underwent a photocycle characterized by the very fast formation of M intermediate. The subsequent intermediate, named P-620, could be assigned to the 0 intermediate in other H+ pumps. In contrast to conventional 0, the accumulation of P620 remains prominent, even at high pH. Flash-induced absorbance changes suggested that there exists only one kind of photocycle at any pH. However, above pH 7, RpActR shows heterogeneity in the H+ transfer sequences: one first captures H+ and then releases it during the formation and decay of P-650, while the other first releases H+ prior to H+ uptake during P-620 formation. (C) 2016 Elsevier B.V. All rights reserved

Modulation of Neurogenesis through the Promotion of Energy Production Activity Is behind the Antidepressant-Like Effect of Colonial Green Alga, Botryococcus braunii

Algae have been recognized as important resources providing functional components due to their capacity to exert beneficial effects on health. Therefore, there is increasing interest in investigating the biological activity of algae. In this study, we evaluated the antidepressant-like effect of the administration of 100 mg/kg/day of the ethanol extract of colonial green alga Botryococcus braunii (EEB) for 14 consecutive days in the forced swimming test (FST)-induced depression in imprinting control region (ICR) mice. Imipramine, a commercial antidepressant drug, was used as a positive control. In addition, we investigated the molecular mechanisms underlying the effect of EEB by measuring ATP production and by assessing any change in gene expression at the end of the treatment using real-time polymerase chain reaction (PCR) and microarray assays. We showed that the immobility time in the water-administered control (FST stress) group gradually increased from day 1 to day 14. However, treatment with EEB caused a significant decrease of immobility time in the FST compared with that in the FST stress group. Microarray and real-time PCR results revealed that EEB treatment induced variation in the expression of several genes associated with neurogenesis, energy metabolism, and dopamine synthesis. Interestingly, we revealed that only EEB treatment enhanced the promotion of energy production, while treatment with imipramine was ineffective. Our study provides the first evidence that B. braunii enhances energy production, which may contribute to the modulation of neurogenesis and to the enhancement of dopaminergic function, in turn potentially underlying the antistress- and antidepressant-like effects that we observed

Preparation and Observation of Fresh-frozen Sections of the Green Fluorescent Protein Transgenic Mouse Head

Hard tissue decalcification can cause variation in the constituent protein characteristics. This paper describes a method of preparating of frozen mouse head sections so as to clearly observe the nature of the constituent proteins. Frozen sections of various green fluorescent protein (GFP) transgenic mouse heads were prepared using the film method developed by Kawamoto and Shimizu. This method made specimen dissection without decalcification possible, wherein GFP was clearly observed in an undamaged state. Conversely, using the same method with decalcification made GFP observation in the transgenic mouse head difficult. This new method is suitable for observing GFP marked cells, enabling us to follow the transplanted GFP marked cells within frozen head sections

Engineering Study of a Pilot Scale Process Plant for Microalgae-Oil Production Utilizing Municipal Wastewater and Flue Gases: Fukushima Pilot Plant

This article presents an engineering study of an integrated system to produce bio-oil from microalgae biomass. The analysis is based on a pilot plant located at Minami-soma Fukushima, Japan, which further simulates 1 ha based-cultivation. Municipal wastewater and flue gases were utilized as nutrient sources for the microalgae culture of the proposed design. A flow sheet diagram of the integrated plant was synthesized by process engineering software to allow simulation of a continuous system. The design and sizing of the process equipment were performed to obtain a realistic estimation of possible production cost. The results demonstrated that nutrient savings was achieved by wastewater and CO2 utilization to the polyculture of native microalgae. Process simulation gave an estimated CO2 sequestration of 82.77 to 140.58 tons ha−1year−1 with 63 to 107 tons ha−1year−1 of potential biomass production. The integrated process significantly improved the energy balance and economics of biofuel production and also the wastewater treatment plant (WWTP). The economic analysis confirmed that higher biomass production and technology improvement were required to achieve operational feasibility and profitability of the current microalgae-based bio-oil production

Giant cell tumor expanded into the thoracic cavity with spinal involvement

This article describes a case of a giant cell tumor that expanded into the thoracic cavity and through the spinal canal into the vertebrae. A 36-year-old man presented with a 6-month history of back pain and dyspnea. Plain chest radiographs showed a huge mass accompanied by right pleural effusion. The mass involved the 12th thoracic spine, and the spinal cord was severely compressed. The tumor was resected with a 2-stage procedure. As a first stage to separate the tumor from the anterior vital structures under direct vision, thoracic surgeons performed a right thoracotomy with chest wall reconstruction from the 8th to 11th ribs. The right lung and inferior vena cava were gently retracted, and the tumor was carefully detached from these structures. We were not able to separate the tumor from the right diaphragm due to severe invasion; therefore, we performed partial resection of the right diaphragm with the tumor. After excision of the anterior part of the tumor, the thoracic wall was reconstructed with the right eighth rib and Marlex mesh. When the patient\u27s general condition improved 2 weeks later, spondylectomy by posterior approach was performed. We achieved excision of a giant cell tumor that had expanded into the thoracic cavity and through the spinal canal into the vertebrae. The patient had achieved full rehabilitation with no neurological or respiratory abnormalities at 7 years postoperatively